Search Results (124)

Background: The 99th-percentile upper reference limit (99% URL) is essential for high-sensitivity cardiac troponin I (hs-cTnI) assays to diagnose acute coronary syndromes. We derived the gender-specific 99% URLs for the new Snibe hs-cTnI assay and verified its high-sensitivity performance. Methods: The Snibe hs-cTnI assay has a claimed limit of blank/detection/quantitation of 0.5/1.0/2.0 ng/L, a precision of 5.67% (@ 9.83 ng/L), 4.65% (@ 21.0 ng/L) and 3.77% (@ 174 ng/L), an analytical measuring range of 1.00–500,000 ng/L, and a claimed 99% URL of 20.1/11.8/17.5 ng/L (males/females/overall). Assay precision was evaluated using kit control materials. A total of 846 (M 444, F 402) anonymized leftover samples from healthy individuals were assessed for the derivation of 99% URLs. Results: The inter-assay CV was 6.2/4.4% (@ 10.1/20.6 ng/L). The 10/20% CV corresponded to a concentration of 5.2/3.0 ng/L. The assay recorded detectable hs-cTnI values in 64% of women and 70% of men. Hs-cTnI values were significantly lower in females than males (females vs. males median 2.6 vs. 3.2 ng/L, Mann–Whitney p = 0.005). The derived 99% URL for the population was 7.7 ng/L (90% CI 6.93–22.8 ng/L) for women and 13.7 ng/L (90% CI 10.6–41.4 ng/L) for men. When confined to subjects ≥40 years old only, males had a 99% URL of 14.8 ng/L (n = 412, 90% CI 10.6–41.4 ng/L) and 8.1 ng/L (n = 377, 90% CI 6.95–22.8 ng/L) for females, respectively. Conclusions: The Snibe hs-cTnI assay fulfills the requirements of a hs-cTn assay. Gender-specific 99% URLs are derived for this assay and they increased with age. Full article

This study characterized extracellular vesicles (EVs) isolated from medicinal herb Centella asiatica tissue culture and investigated their therapeutic properties using in vitro assays and a ultraviolet (UV)-induced damage mouse model. EVs were isolated from C. asiatica tissue culture and characterized by nanoparticle tracking analysis, and cytotoxicity, antioxidant, anti-melanin, and anti-inflammation properties were evaluated by in vitro assays. C. asiatica EVs were found to contain high levels of polyphenols and mitigate hydrogen peroxide-induced intracellular reactive oxygen species (ROS). The EVs were further able to reduce intracellular melanin content and tyrosinase activity. They exhibited anti-inflammatory effects by downregulating the expression of pro-inflammatory genes, COX2, as well as nitric oxide production. In the UV-induced photodamage mouse model, gels with or without EVs were applied to the UV-damaged site, skin appearance was observed daily, and skin histopathology was analyzed on day 7. In mice with UV-induced skin damage, the daily application of C. asiatica EV gel reduced skin epidermis thickness and inflammation compared to UV-only or blank gel at seven days after UV irradiation. The beneficial effects of C. asiatica EVs on skin quality warrant further studies as promising agents in skin care applications. Full article

The impact of encapsulating gemcitabine (GEM) into nanoparticles on its delivery remains underexplored, with the potential benefits of targeted drug delivery and stimuli-responsive release yet to be fully clarified. Herein, we designed a near-infrared (NIR) light-responsive polymeric nanoparticle, ZnPc@P(PEG-CMA-TKGEM), which integrates reactive oxygen species (ROS) generation and cell imaging capabilities. The self-assembled polymeric micelles exhibit a hydrodynamic size of ~134 nm. Under NIR irradiation, the cumulative drug release rate reaches 51% within 48 h, which is three times higher than that of the non-irradiated control group. In cytotoxicity assays, the cell viability of the NIR-irradiated drug-loaded group is approximately 17%, while that of the NIR-irradiated blank group (without drug loading) remains above 80%. These results confirm that the nanocarriers successfully deliver GEM to target cells and achieve controlled drug release via NIR stimulation. Full article

Background: We evaluated the Sysmex Highly Integrated Single-Cartridge Luminescence Immunoassay System (HISCL) hs-cTnT assay, and compared its performance to the Roche assay, with derivation of 99th-percentile upper reference limits (99% URLs) for healthy subjects. We assessed the effect of increasing age/decreasing eGFR on the HISCL hs-cTnT. Methods: We verified assay limits of blank/detection, precision and the functional sensitivity. Samples were analyzed on both the Sysmex HISCL and Roche Elecsys analyzers for method comparison. Results: The HISCL assay limit of blank/detection was 1.3/1.9 ng/L, and concentrations corresponding to 20/10% CVs were 1.8/3.3 ng/L. Assay precision of kit controls at 3253 ng/L was 2.2% and at 106 ng/L was 2.5%. Linear regression analysis (n = 2151) showed good agreement (r = 0.95) with the Roche hs-cTnT. Bland–Altman (Roche/HISCL) analysis for samples with hs-cTnT ≤ 52 ng/L showed a mean absolute difference of 3.5 ng/L; for hs-cTnT > 52 ng/L, the mean difference was 2.8%. In a cardio-renal healthy population (n = 1004), the 99% URLs were 14.4/17.0/13.9 ng/L for overall/male/female, respectively; assay CV% was below 10% at these levels. More than 50% of the hs-cTnT in the healthy male and female subjects were measurable above the limit of detection. Hs-cTnT increased with increasing age and decreasing eGFR. Conclusions: In conclusion, the Sysmex HISCL hs-cTnT fulfils the criteria for a high-sensitivity assay, with specific 99th URLs for males and females. Expectedly, the baseline Sysmex hs-cTnT increases with age and decreasing eGFR. Full article

Background: We investigated the role of a high-sensitive cardiac troponin T (hsTnT) increase below the upper limit of normal (ULN) in patients with breast cancer (BC). hsTnT assays accurately quantify very low plasma troponin concentrations and enable the early detection of cardiomyocyte injury before a drop in the left ventricular ejection fraction (LVEF). The increase in hsTnT below the ULN in response to chemotherapy has not previously been studied. Method: This was an open-label pilot study. Female patients with newly diagnosed BC scheduled to receive systemic cancer treatment were recruited. Blood sampling and echocardiography were performed at baseline, at 3 and 6 months of cancer treatment. hsTnT concentrations were measured using the Elecsys TnT hs assay (Roche Diagnostics). The limit of blank and 99th percentile cutoff values for the hsTnT assay were 3 and 14 ng/L. We calculated the rise in hsTnT (ΔhsTnT) by the difference (%) between its baseline level and during follow-up (FU) in each patient. Results: Among eligible subjects, we excluded 4 patients before the start of treatment and 17 patients during the follow-up with values for the hsTnT >14 ng/L. Finally, 60 women with a median age of 48.6 ± 1.3 years were included in the study. The median baseline hsTnT concentration was 5.5 ± 1.4 ng/L. During 6 months of cancer treatment, hsTnT increased in all patients by up to 10–305% from baseline, with an average of 94.2%. LV EF was normal at baseline and decreased significantly compared to the value before cancer treatment (61.9 ± 3.3% vs. 56.3 ± 7.0%; p < 0.045). We correlated the hsTnT rise with a drop in LV EF ≥ 10% from its baseline level. Logistic regression analysis showed that Δ hsTnT has a good predictive value for LV dysfunction, 0.78 (p = 0.05), 95% CI (0.67–0.90). The increase in hsTnT > 81% was determined as the optimal threshold value for detecting early biochemical cardiotoxicity. Conclusion: It was investigated that hsTnT rise within the cutoff < 14 ng/L can be used as a marker of early biochemical cardiotoxicity and is valuable for predicting LV drop in 6 months of FU. We conclude that BC patients with increased hsTnT plasma concentration > 81% from the baseline value should be considered as high-risk patients for cardiotoxicity and need more precise cardiac monitoring and early preventive medical intervention strategies. Full article

CSNPs synthesized via the ionic gelation method have emerged as a promising nanoplatform in diverse fields such as pharmaceuticals, nanotechnology, and polymer science due to their biocompatibility, ease of fabrication, and tunable properties. This study focuses on the development and characterization of LL37-loaded CSNPs, designed to enhance antibacterial efficacy while maintaining biocompatibility. This study pioneers a systematic loading optimization approach by evaluating the encapsulation efficiency (%EE) of antimicrobial peptide LL37 across multiple concentrations (7.5, 15, and 30 µg/mL), thereby identifying the formulation that maximizes peptide incorporation while preserving controlled release characteristics. The multi-concentration analysis establishes a new methodological benchmark for peptide delivery system development. To achieve this, CSNPs were optimized for size and stability by adjusting parameters such as the chitosan concentration, pH, and stabilizer. LL37, a potent antimicrobial peptide, was successfully encapsulated into CSNPs at concentrations of 7.5, 15, and 30 µg/mL, yielding formulations with favorable physicochemical properties. Dynamic light scattering (DLS) and Zeta sizer analyses revealed that blank CSNPs exhibited an average particle size of 180.40 ± 2.16 nm, a zeta potential (ZP) of +40.57 ± 1.82 mV, and a polydispersity index (PDI) of 0.289. In contrast, 15-LL37-CSNPs demonstrated an increased size of 210.9 ± 2.59 nm with an enhanced zeta potential of +51.21 ± 0.93 mV, indicating an improved stability and interaction potential. Field emission scanning electron microscopy (FE-SEM) analyses exhibited the round shaped morphology of nanoparticles. The release profile of LL37 exhibited a concentration-dependent rate and showed the best fit with the first-order kinetic model. Cytocompatibility assessments using the XTT assay confirmed that both blank and LL37-loaded CSNPs did not exhibit cytotoxicity on keratinocyte cells across a range of concentrations (150 µg/mL to 0.29 µg/mL). Notably, LL37-loaded CSNPs demonstrated significant antibacterial activity against E. coli and S. aureus, with the 15-LL37-CSNP formulation exhibiting superior efficacy. Overall, these findings highlight the potential of LL37-CSNPs as a versatile antibacterial delivery system with applications in drug delivery, wound healing, and tissue engineering. Full article

Applications of lanthanide chemistry have been successful in metallics and the petroleum industry. In the medical realm, lanthanides have shown utility in radiotherapy agents, photodynamic therapy agents, and magnetic resonance imaging (MRI) contrast agents. The lanthanide group elements have a few known biological roles, notably among some bacteria and the yeast Saccharomyces cerevisiae, which have been used as models for changes in gene expression. However, the systematic effects of lanthanide nitrates on eukaryotic cell model systems have not yet been reported. This study presents the first documented effects on cell viability, after acute incubations of various lanthanide nitrate salts, using axenic C6 glial or PC12 neuronal cells in vitro. Cultures were exposed to a 1 mM concentration of lanthanide nitrate salts for 24 h. In comparison to the saline control, several cultures demonstrated significantly lower cell viability, as measured by the MTT viability assay. Data were analyzed as an average absorbance of n = 4 replicate samples, corrected for the average absorbance of cell-free blanks. The reported results were normalized to the average of the saline control cells. Among the 13 lanthanides tested, Praseodymium, Holmium, Erbium, Thulium, and Ytterbium nitrates exhibited the most pronounced inhibitory effects, resulting in over 40% reduction in cell viability at 1 mM for either or both cell types. Recovery after lanthanide exposure also was cell-type-dependent as well as lanthanide-type-dependent, with Lutetium having the greatest effect on both cell types. PC12 cells displayed greater sensitivity for inhibition than the C6 cells with some of the lanthanides but not all. Furthermore, the controls of sodium nitrate and calcium nitrate showed only modest discernible impacts on cell viability for PC12 and C6 cells, highlighting the role of the lanthanides in influencing cell viability. Full article

Background: Metritis, a common postpartum uterine infection in bovines, poses substantial challenges in livestock management, including compromised fertility and economic losses. Poor uterine drug penetration and systemic side effects, necessitating innovative localised delivery systems and limiting current systemic antibiotic treatments. Aim: This study aimed to develop and evaluate the potential effect of the ofloxacin-loaded hydrogel as a localised drug delivery system to treat metritis in bovine. The focus was on achieving sustained drug release, enhanced antibacterial efficacy and reduced inflammation in the endometrium. Materials and Methods: The CS/PVA hydrogel was synthesised using a freeze–thaw method and further optimised for drug encapsulation efficiency (96.7 ± 2.1%), stability and biocompatibility. Physicochemical characterisation included swelling behaviour, mechanical properties and rheological analysis. In vitro drug release profiles in the simulated uterine fluid were assessed over 72 h and antibacterial activity was tested against common uterine pathogens such as Escherichia coli and S. aureus. In vivo studies were conducted on bovines diagnosed with endometritis to evaluate clinical recovery. Results: The SEM image of the ofloxacin-loaded CS/PVA hydrogel resulted in a smooth and porous structure demonstrating larger pore size than the blank. The rheological study suggested higher stability and elastic behaviour. Antibacterial assays on E. coli and S. aureus revealed significant inhibition zones, respectively, indicating potent efficacy. In vivo, evaluated on treated bovine, reduced bacterial loads were exhibited (2.86 × 10^5A CFU/mL → 6.37 × 10^2B CFU/mL), clinical improvement was marked and uterine inflammation was resolved. Conclusions: Ofloxacin-loaded hydrogels represent a promising localised treatment for bovine metritis, offering sustained antibacterial action and improved clinical outcomes. This approach addresses the limitations of systemic antibiotic therapies and provides a practical solution for enhanced veterinary care. Further studies are recommended to validate these findings in more extensive field trials and explore commercialisation potential. Full article

Aflatoxin M1 (AFM1) appears in the milk of animals that have consumed feed contaminated with aflatoxin B1. AFM1 presence in milk is regulated by the European Commission, which has set the maximum allowable limits for adult and infant consumption to 50 and 25 pg/mL, respectively. Here, a rapid and sensitive method for detecting AFM1 in milk based on an immersible silicon photonic chip is presented. The chip features two U-shaped silicon nitride waveguides formed as Mach–Zehnder interferometers. One interferometer is functionalized with AFM1–bovine serum albumin conjugate and the other with BSA to serve as a blank. The chip is connected to a broad-band white LED and a spectrophotometer by a bifurcated optical fiber and an assay is performed by immersing the chip in a mixture of milk with the anti-AFM1 antibody. Then, the chip is sequentially immersed in biotinylated anti-rabbit IgG antibody and streptavidin solutions for signal enhancement. The assay is completed in 20 min and the detection limit for AFM1 in undiluted milk is 20 pg/mL. Given its analytical performance and the absence of pumps and fluidics that lead to a compact instrument design, the proposed immunosensor is ideal for the on-site detection of AFM1 in milk samples. Full article

Background/Objectives: The NavDx+Gyn blood test detects and quantifies fourteen HPV types in various sample types to provide a reliable means of detecting and monitoring HPV-driven gynecologic cancers. NavDx+Gyn is an extension of the NavDx assay, which identifies five high-risk HPV types. NavDx has been clinically validated in multiple independent studies for the surveillance of HPV-driven oropharyngeal cancer and has been integrated into clinical practice by over 1300 healthcare providers at over 500 medical sites in the US. The NavDx+Gyn assay incorporates an analysis of nine additional high-risk HPV types. Here, we report a detailed analytical validation of the NavDx+Gyn assay for use in cervical, vaginal, and vulvar cancer patients to detect fourteen high-risk HPV types related to HPV-driven gynecologic cancers. Methods: Parameters include specificity as measured by limits of blank (LoBs) and sensitivity illustrated via limits of detection and quantitation (LoDs and LoQs). Results: The LoBs were between 0 and 0.0926 copies/μL, LoDs were 0.1009 to 0.3147 copies/μL, and LoQs were 0.1009 to 0.3147 copies/μL, demonstrating the high analytic sensitivity and specificity provided by NavDx+Gyn. In-depth evaluations, including accuracy and intra- and inter-assay precision studies, were shown to be within acceptable ranges. Regression analysis revealed a high degree of correlation between expected and effective concentrations, demonstrating excellent linearity (R² > 0.99) across a broad range of analyte concentrations. Conclusions: These results demonstrate that NavDx+Gyn accurately and reproducibly detects fourteen types of high-risk HPV, which aids in the diagnosis and surveillance of the vast majority of HPV-driven gynecologic cancers. Full article

Background/Objectives: Atorvastatin (ATR), an antihyperlipidemic drug with a potential antibacterial effect, was investigated in this study. Like other statins, ATR has been repurposed for several uses, ranging from anti-inflammatory to antimicrobial applications, and has demonstrated successful results. However, the efficacy of ATR is limited by its low solubility, indicating an opportunity for its encapsulation in a nanotechnology-based drug delivery system. Methods: Nanostructured lipid carrier (NLC) formulations were prepared using high-pressure homogenization and ultrasonication. The formulations were characterized, including their particle size, polydispersity index, zeta potential, encapsulation efficiency, and in vitro release. Antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli (E. coli), and Staphylococcus aureus (S. aureus) was evaluated using the growth curve (bacterial growth over time) and well diffusion methods (zone of inhibition and minimum inhibitory concentration (MIC) determination). The crystal violet assay was employed to assess biofilm inhibition. Results: The NLC formulations were optimized, and the size and zeta potential of the blank nanoparticles were 130 ± 8.39 nm and −35 ± 0.5 mV, respectively. In comparison, the encapsulated NLCs had a size of 142 ± 52.20 nm and a zeta potential of −31 ± 1.41 mV. The average encapsulation efficiency was 94%, and 70% of the drug was released after 24 h. The ATR-loaded NLCs showed significantly enhanced antibacterial activity by reducing the minimum inhibitory concentration by 2.5-fold for E. coli, 1.8-fold for S. aureus, and 1.4-fold for MRSA, and promoting more effective bacterial growth inhibition. Notably, biofilm inhibition was significantly improved with ATR-NLCs, achieving 80% inhibition for S. aureus, 40% for E. coli, and 30% for MRSA, compared to free ATR (p < 0.001). These findings suggest that NLC encapsulation enhances ATR’s antimicrobial efficacy and biofilm suppression. Conclusions: This study identified NLCs as successful carriers of ATR, significantly enhancing its antibacterial efficacy and biofilm inhibition capabilities. This formulation, which shows antimicrobial potential against both Gram-positive and Gram-negative bacteria, should be further studied and developed against different resistant microbial strains. Full article

The immunohistochemical (IHC) or fluorescence/chromogenic in situ hybridization (FISH/CISH) assays for assessing HER2 are now recommended by the American Society of Clinical Oncologists and the College of American Pathologists, but there are an increasing number of published studies describing alternative diagnoses at the molecular level. Inspired by these studies, we established a laboratory-developed test (LDT) to analyze HER2 status not only at the gene expression level but also at the gene copy number. A precise copy number calculation was fulfilled including the Control Genomic DNA of known concentration, which allowed subsequent assay validation at the DNA level. The results were reported according to the concordant results of the DNA and RNA approaches. By comparing with IHC determination, completely identical results were found in ten blank samples, which underlines the legitimacy of molecular biological approaches in this diagnostic field. An equivocal sample that was positive by IHC and qPCR was found to be negative by the FISH and so it may change the choice of personalized medicine. The topic of this short communication will hopefully contribute to allowing IVD-certified diagnostics based on the HER2 gene expression profile or copy number to be tested in the Czech Republic as well. Full article

This study aimed to explore the therapeutic potential and mechanisms of Artemisia argyi essential oil (AAEO) in managing feline herpesvirus type 1 (FeHV-1) infections. FeHV-1, the causative agent of feline viral rhinotracheitis (FVR), leads to severe respiratory and systemic complications in cats. In this study, 35 cats were divided into blank, FeHV-1 infection, and AAEO treatment groups (high, medium, and low doses). In vivo experiments demonstrated that AAEO alleviated clinical symptoms, reduced tissue damage, and modulated immune responses. The AAEO-treated groups showed higher survival rates, stabilized body temperatures, and less severe weight loss compared to the FeHV-1 group. Histopathological analysis revealed improved integrity in nasal, tracheal, and bronchial tissues. Transcriptomic and proteomic analyses identified critical pathways, such as IL-17 signaling, influenced by AAEO treatment, highlighting its role in suppressing inflammation and protecting tissue integrity. In vitro assays revealed that AAEO has concentration-dependent cytotoxicity in feline kidney cells (F81) and provides protective effects when used as a pre-treatment. These findings suggest that AAEO enhances host immune defenses and mitigates FeHV-1-induced damage through immune modulation and tissue protection. Full article

The present work reports the development, optimization, and validation, of a methodology to determine lidocaine, procaine, tetracaine, and benzocaine in urine matrices. Two extractive preconcentration techniques, solid-phase microextraction (SPME) LC Tips and bar adsorptive microextraction (BAμE), were studied and applied to the four target anesthetics, followed by gas chromatography-mass spectrometry (GC-MS) analysis. Several parameters that could affect microextraction and back-extraction were optimized using two different designs of experiments (Box–Behnken and full-factorial) to maximize extraction efficiency from aqueous media. Under optimized experimental conditions, the BAμE technique showed better performance than SPME LC Tips and was chosen for validation assays and urine sample analysis. In blank urine, the BAµE/GC-MS methodology revealed suitable sensitivity (LOD between 2 and 18 ng/mL), good linearity (r² ≥ 0.9945) between 0.5 and 30.0 µg/mL and recovery yields of 30.3–97.9%. Good precision (%RSD ≤ 8.8%) and accuracy (bias % between −15.9 and 15.0%) values were achieved. The developed methodology was successfully applied to the target anesthetics analysis of volunteers’ urine matrices and proved to be an environmentally friendly alternative to monitor trace levels of local anesthetics in complex matrices compared to other extraction techniques. Full article

Chronic wounds represent a persistent clinical challenge due to prolonged inflammation and impaired tissue repair mechanisms. Cannabidiol (CBD), recognized for its anti-inflammatory and pro-healing properties, shows therapeutic promise in wound care. However, its delivery via lipid nanoparticles (LNPs) remains challenging due to CBD’s inherent instability and low bioavailability. This study developed and characterized a novel hydrogel scaffold composed of CBD-loaded LNPs (CBD/LNPs) integrated into a polyvinyl alcohol (PVA) and sodium alginate (SA) matrix, designed to enhance wound repair and mitigate inflammation. The characteristics of the hydrogel scaffold were observed including the degree of swelling and LNPs’ release profiles. Furthermore, in the results, CBD/LNPs displayed enhanced stability and reduced cytotoxicity compared to unencapsulated CBD. In vitro assays demonstrated that CBD/LNPs significantly promoted fibroblast migration in gap-closure wound models and reduced intracellular reactive oxygen species, supporting their potential as a biocompatible and efficacious agent for cellular repair and oxidative stress attenuation. In vivo experiments using adult male Wistar rats with aseptic cutaneous wounds revealed that treatment with CBD/LNP-PVA/SA hydrogel scaffold significantly accelerated wound closure relative to blank hydrogel controls, demonstrating a substantial reduction in the wound area over time. Histological analysis confirms notable improvements in skin morphology in wounds treated with CBD/LNP-PVA/SA hydrogel scaffold with evidence of accelerated epithelialization, enhanced collagen deposition, and increased dermal thickness and vascularization. Additionally, skin histology showed a more organized epidermal layer and reduced inflammatory cell infiltration in CBD/LNP-PVA/SA hydrogel scaffold-treated wounds, corresponding to a 35% increase in the wound closure rate by day 28 post-treatment. These findings suggest that CBD/LNP-PVA/SA hydrogel scaffolds facilitate inflammation resolution and structural wound healing through localized, sustained CBD delivery. The dual anti-inflammatory and wound-healing effects position CBD/LNP-PVA/SA hydrogel scaffold as a promising approach for chronic wound management. Future investigations are warranted to elucidate the mechanistic pathways by which CBD modulates the skin architecture and to explore its translational applications in clinical wound care. Full article